Product Catalog. biolegend.com General Information. Technical Charts and Protocols. Mouse Immunology Antibodies

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1 General Information Technical Charts and Protocols Mouse Immunology Antibodies 2014 Product Catalog Rat Immunology Antibodies Human Immunology Antibodies Cytokine and Chemokine Antibodies Recombinant Proteins for Bioassay Immunoassays Cell Biology Antibodies Secondary Reagents 2 Isotype Controls Buffers, Solutions, and Chemicals Index of Catalog Numbers Master Index 1

2 Table of Contents Catalog Abbreviations Guide...3 Company Profile...4 Ordering Information...5 Custom Solutions Team...6 Technical Services...7 Quality and Satisfaction Guarantee...7 Collaborate with Us...8 Terms and Conditions...8 Product Warranties and Use Limitations...9 Trademarks...9 Patents...9 Technical Charts and Protocols Brilliant Violet Fluorophores...12 Fluorophores for Flow Cytometry...14 Human CD Molecules, HLDA9 Workshop...16 Biofunctional Antibodies - LEAF and Ultra-LEAF...30 Cross-Reactivity Chart, Non-Human Primate...40 Cross-Reactivity Chart, Other Species...43 TNF Receptor and Ligand Superfamilies...45 Cell Surface Immunofluorescence Staining Protocol...47 Intracellular Cytokine Staining Protocol...48 FOXP3 Intracellular Staining Protocol...49 Th17 Polarization of Mouse CD4 Cells...49 Western Blotting Protocol...50 Immunoprecipitation Protocol...51 General Sandwich ELISA Protocol...52 ELISPOT Protocol...54 Immunofluorescence Protocol...55 Immunocytochemistry and Immunohistochemistry...56 Mouse Immunology Antibodies LEGENDScreen...58 Mouse Immunophenotyping...59 Rat Immunology Antibodies Human Immunology Antibodies LEGENDScreen Human Immunophenotyping Cytokine and Chemokine Antibodies Chemokines and their Receptors Cytokine/Chemokine Intracellular Staining Guide Cytokine Neutralization with Antibodies Mouse Cytokine and Chemokine Antibodies Rat Cytokine and Chemokine Antibodies Chicken Antibodies Human Cytokine and Chemokine Antibodies Recombinant Proteins for Bioassay Immunoassays ELISA Kits and Sets ELISA/ELISPOT Antibody Pairs and Standards Flow Cytometry Kits and Cocktails Cell Biology Antibodies Secondary Reagents Isotype Controls Buffers, Solutions, and Chemicals Zombie Aqua / Zombie NIR /Zombie Yellow Index of Catalog Numbers Master Index Toll Free BIOLEGEND ( )

3 For your convenience, here is a list of abbreviations used throughout this catalog: aa:...amino acids Ab: Antibody Activ:...Activation AKP: Alkaline phosphatase APC: Allophycocyanin Apop: Apoptosis Ar Ham...Armenian Hamster Av:...Avidin BA: Bioassay BCA:...Bicinchoninic acid protein assay Block:...Blocking Bov...Bovine (Cow) BV421 : Brilliant Violet 421 BV510 : Brilliant Violet 510 BV605 : Brilliant Violet 605 BV650 : Brilliant Violet 650 BV711 :...Brilliant Violet 711 BV785 : Brilliant Violet 785 CMCD:...Complement-mediated cell depletion Chick...Chicken ChIP: Chromatin Immunoprecipitation CNS: Central nervous system Costim:...Costimulatory or costimulation Cy:...Cyanine (dye) Cyt: Cytotoxicity CyTOF : Mass Cytometry Deplete or Depletion:...Cell depletion ELISA:...Enzyme-linked immunosorbent assay ELISA Capture/Detection: Capture or detection reagent for enzyme-linked immunosorbent assay ELISPOT:...Enzyme-linked immunosorbent spot assay ELISPOT Capture/Detection:...Capture or detection reagent for enzyme-linked immunosorbent spot assay EU:...Endotoxin Unit FA:...Functional assay FC: Flow cytometry FITC: Fluorescein isothiocyanate GPI: Glycosylphosphatidylinositol Catalog Abbreviations Guide G Pig...Guinea Pig Gt...Goat HRP:...Horseradish peroxidase Hs...Horse (Equine) Hu...Human ICC: Immunocytochemistry ICFC: Intracellular Staining for Flow Cytometry IF: Immunofluorescence microscopy Ig:...Immunoglobulin IHC:...Immunohistochemistry IP:...Immunoprecipitation kd:...kilodalton LEAF :...Low Endotoxin, Azide-Free (<0.1 EU/µg of protein) LPS:...Lipopolysaccharide µg:...microgram mab:...monoclonal antibody mg:...milligram Ms...Mouse MHC:...Major histocompatibility complex Neut:...Neutralization ng:...nanogram NHP...Non-Human Primate NK:...Natural killer cells PBMC:...Peripheral blood mononuclear cells PE (or R-PE):...Phycoerythrin pg:...picogram Rab...Rabbit RIA: Radioimmunoassay SAv: Streptavidin SDS-PAGE:...SDS polyacrylamide gel electrophoresis Stim:...Stimulation Syr Ham...Syrian Hamster TCR:... T cell receptor TNFRSF:...TNF receptor superfamily TNFSF:...TNF superfamily Treg:...T regulatory cells Ultra-LEAF :...Lower Endotoxin, Azide-Free (<0.01 EU/µg of protein) WB:...Western blotting 3

4 Company Profile BioLegend develops world-class, cutting-edge antibodies and reagents for biomedical research, manufactured in our state-of-the-art facility in San Diego, CA. Our mission is to accelerate research and discovery by providing the highest quality products at an outstanding value, along with superior customer service and technical support. Our product areas include cell immunophenotyping, cytokines and chemokines, adhesion, cancer research, T regulatory cells, stem cells, innate immunity, cell-cycle analysis, apoptosis, and modificationspecific antibodies. All of BioLegend s reagents are supported by superior customer service and come with a quality guarantee. Our aggressive product development program, through technology licensing, collaborations, and internal hybridoma development, produces strategic reagents for use in a variety of applications including: Flow cytometry and mass cytometry ELISA and ELISPOT Immunoprecipitation and ChIP Western blotting Immunofluorescence microscopy Immunohistochemistry In vitro and in vivo functional assays See inside back cover for complete contact information. USA, San Diego Headquarters biolegend.com Japan biolegend.com/jp Germany biolegend.com/de United Kingdom biolegend.com/uk BioLegend European Distribution Center Phone: +44(0) Fax: +44(0) BioLegend offers the broadest selection of fluorochrome conjugates for multi-color flow cytometry. With our Custom Solutions Team, BioLegend is well positioned to offer a wide range of custom products and services tailored to our customers specific needs. Incorporated in June 2002, BioLegend is located in San Diego, in the midst of one of the nation s largest and fastest growing biotechnology clusters. Quality Management System BioLegend in San Diego, CA is now operating under an ISO 9001:2008 and ISO 13485:2003 certified quality management system. This certifies an effective quality management system, based on documentation and continuous improvement, that meets international standards for medical device and reagent manufacturing. 4 Toll Free BIOLEGEND ( )

5 Ordering from US and Canada: Our Customer Service Department is available 7:00 am to 5:00 pm (PST), Monday through Friday to take and process your orders. On-line: Visit biolegend.com/ordering Phone/Fax: Phone (Toll-Free US & Canada): BioLegend ( ) Phone (International): Fax: For most efficient order processing, please be sure that the following information is included on your purchase order or correspondence: 1. Name of institution or company 2. Account number (if available) 3. Shipping address 4. Billing address 5. Catalog number, description, and correct price 6. Quantity and size required 7. Name and phone number of purchasing agent 8. Name and phone number of the end user 9. Purchase Order number 10. BioLegend quotation number (if applicable) Shipping from BioLegend Headquarters: All orders are shipped F.O.B. San Diego, California. Shipping and handling charges will be prepaid and added to the invoice. Domestic orders will normally be shipped by overnight air express. BioLegend reserves the right to select the best packaging and shipping method which will ensure the stability of the product and allow efficient order tracking. Our reagents are stable when shipped at ambient temperature. Please store our products under the conditions indicated on the technical information sheet upon arrival. Domestic shipments are sent overnight Monday through Thursday. International shipments are typically sent on Fridays and Mondays. Ordering from Europe See Inside Back Cover, BioLegend Worldwide, for complete contact Information. Germany: Visit biolegend.com/de and click Kontakt, Ordering for ordering instructions. Ordering Information United Kingdom: Visit biolegend.com/uk and click Contact, Ordering for ordering instructions. Italy: Visit biolegend.com/it and click Contact, Ordering for ordering instructions. Europe, Other Countries: Contact BioLegend Europe to place an order or be directed to our distributor. Inquiries: Technical Support: Phone: +44(0) Fax: +44(0) Shipping from the BioLegend European Distribution Center: The BioLegend European Distribution Center in the United Kingdom maintains the majority of our catalog items in stock for same-day delivery. We will ship via our preferred courier and shipping charges vary, depending on the weight of the shipment and the destination country. Customers will be responsible for payment of any applicable duties and taxes upon shipment arrival in their respective country. Prepayment is required. Ordering from Japan See Inside Back Cover, BioLegend Worldwide, for complete contact Information. Japan: Visit biolegend.com/jp and click お 問 い 合 わせ, Ordering for ordering instructions. Ordering from All Other Countries Customers located outside of the United States may order through BioLegend distributors or through BioLegend directly. We have extensive experience in shipping to many countries directly. We will ship via our preferred courier and shipping charges vary, depending on the weight of the shipment and the destination country. Customers are responsible for payment of duties and taxes upon shipment arrival in their respective country. Orders from outside the USA and Canada typically require prepayment before shipping. Direct ordering instructions are the same as Ordering from the US and Canada, but include your Tax ID number for customs, as well as a copy of the import permit/license (if applicable). International shipments are typically sent on Fridays and Mondays. International Distributors: Visit biolegend.com/distributors for complete list of Worldwide Distributors. 5

6 Custom Solutions Team Specialized Products and Services Tailored for Your Research BioLegend s Custom Solutions Team has decades of combined research and manufacturing experience in developing conjugated antibodies, flow cytometry antibody cocktails, ELISAs, multiplex assays, bioassays, recombinant proteins, and custom antibody production. We will collaborate with you to fully understand your needs, develop a specialized custom solution, and then deliver the highest caliber products and services, along with our quality guarantee. Save time and money by leveraging our expertise. Custom Products Custom antibody cocktails in bulk, including cell surface and intracellular targets Lyophilized antibody cocktails Custom antibody conjugation Multiplex immunoassays on multiple platforms ELISA and ELISPOT kits and sets Antibody screening plates Recombinant protein expression Custom Services Biomarker assay development services Sample analysis for biomarkers Custom manufacturing and packaging Monoclonal and polyclonal antibody production Bioassay development Additional needs? Let s discuss. Outstanding Value As with all of our products, we provide excellent and reasonable pricing for our custom solutions. Our products and services are backed by our high quality standards, including: Pre- and post-bottling testing Strict documentation and backup of protocols and data Use of regularly calibrated instruments and equipment Lot-to-lot consistency verified by well-defined specifications Highly-skilled personnel with certified biohazard training Daily monitoring of storage refrigerators and freezers Custom Flexibility We understand that each customer has unique project requirements. That s why we are completely flexible with custom manufacturing, packaging, labeling, bulk sizing, batch delivery timelines, etc. We are here to meet your needs. Let s get started on your custom solution us at CST@biolegend.com, call , or contact your local BioLegend representative. 6 Toll Free BIOLEGEND ( )

7 Technical Services At BioLegend, our Technical Services team is dedicated and trained to answer your questions on our products and their related applications. All of our technical service staff are highly qualified and experienced to address your needs. We welcome any suggestions or comments to help us achieve the highest level of service. For technical support, please call us at (Toll-free US & Canada) or (International), fax your inquiry to , or send your to: Our Technical Services Department hours are 7:00 am to 5:00 pm, Pacific Standard Time, Monday through Friday. Quality and Satisfaction Guarantee Our strict quality control guidelines allow us to offer a 100% satisfaction guarantee on all our products. As exceptional quality and batch-to-batch consistency are of paramount importance for all of us at BioLegend, each batch must pass stringent biochemical and biological/immunological testing requirements. If, however, any product does not perform as indicated on its Technical Data Sheet, we will either immediately replace it or credit you the original purchase price. Please contact our Technical Services Department if you have any questions or concerns about your product. Our Technical Service Scientists are available to help with assay design, suggest relevant positive controls, and offer troubleshooting tips. Your satisfaction is our number one priority. Quality Policy Statement BioLegend is a global leader in bioscience committed to world-class quality, superior customer support, and outstanding value. We achieve this position through scientific innovation, continuous improvement, customer satisfaction, and operating under an effective Quality Management System that meets Regulatory Requirements of ISO 9001:2008 and ISO 13485:2003 7

8 Collaborate with Us BioLegend is committed to providing researchers with the most comprehensive and cutting-edge high quality reagents for life science research. If you have developed reagents or technologies that you feel may be of interest to the life science research community, we would like the opportunity to collaborate with you to commercialize your inventions. We look forward to developing a mutually beneficial collaboration with you. For more information about collaboration and our technology transfer process, please busdev@biolegend.com. Terms and Conditions Pricing Prices reflect list price and are subject to change without notice. Payment Methods and Terms We accept credit card purchases for US and Canada orders; please have your VISA, MasterCard, American Express, or Discover Card account number and expiration date available when placing an order. Purchase order numbers are required for all billed shipments. Payment terms are net 30 days. A 1.5% monthly late charge may be charged to accounts that are over 30 days past due. Orders from outside the USA and Canada typically require prepayment before shipping. Limited Use Label License The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer this product its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for commercial purposes. The buyer may transfer information or materials made through the use of this product to a scientific collaborator, provided that such transfer is not for any Commercial Purpose, and that such collaborator agrees in writing not to transfer such materials to any third party and to use such transferred materials and/or information solely for research and not for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: use of the product or its components in manufacturing; use of the product or its components to provide a service, information, or data; use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or resale of the product or its components, whether or not such product or its components are resold for use in research If the purchaser is not willing to accept the limitations of this limited use statement, BioLegend requires immediate return of the product with a full refund. Order Confirmation and Duplication Written confirmation of a telephone or fax order is not required. However, if confirmation of an order is sent, it should be clearly marked CONFIRMING ORDER DO NOT DUPLICATE. Duplicated shipments due to incorrectly marked confirming orders will be subject to a Restocking Fee of 20% of the order or minimum of $25, whichever is greater, plus shipping charges. Returns A return authorization number is required on all returns. Please contact BioLegend Customer Service Department to obtain the authorization prior to returning any item. All claims for damaged merchandise during transit must be reported immediately to the shipping carrier, and then to BioLegend. Return authorization requests must be made within 10 working days of order receipt. All claims are subject to investigation before return authorization, replacement or credit can be issued. If BioLegend makes a shipping error, we will replace the product in error or the missing product immediately at no charge. If the customer makes an error on the order and is reported to BioLegend within 10 working days, the customer may obtain return authorization and return the product(s) for partial credit. Upon receipt of authorized returned goods, a credit will be issued, less a 20% restocking fee or minimum of $25, whichever is greater, of the product(s) returned and the original shipping and handling charge. To obtain partial credit, product(s) must be returned within 30 days of receiving return authorization. 8 Toll Free BIOLEGEND ( )

9 Product Warranties and Use Limitations Limited Warranty and Liability Please review specifications of the product as published in the product details section of our website or call and request a copy from our Technical Service Department before ordering. Product details on the website generally reflect the applications and conditions for use. Products should be used in accordance with the technical information sheet shipped with the product. BioLegend reserves the right to change product specifications without prior notice. All products supplied by BioLegend are warranted, to the original purchaser only, to conform to our specifications provided with the shipment. This warranty does not extend to any product that has been modified in any way by person(s) other than BioLegend personnel or to any products that are handled contrary to the technical information sheet. Should any product fail to perform as warranted, upon notification to our Technical Service Department and investigation, BioLegend shall promptly replace it at no charge. Our obligation and your only remedy is limited to such replacement of the product or a full credit of the original purchase price of the product. BioLegend reserves the right to discontinue sales of any products or to change product descriptions and formulations at any time without prior notice to purchasers. BioLegend does not make any other warranty or representation whatsoever, whether express or implied, with respect to these products. BioLegend does not make any warranty of suitability, non-patent-infringement, merchantability or fitness for any particular research use or other purposes of any product. The entire risk as to the performance of this product is assumed by buyer. In no event shall BioLegend be liable for incidental, consequential or punitive damages. Product Use Limitations Unless otherwise clearly indicated on product labels, BioLegend products are: FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE Trademarks BioLegend, BioLegend Logo, LEAF, Ultra-LEAF, The path to legendary discovery, TruStain FcX, FluoroFix, Cyto-Last, LEGENDScreen, ELISA MAX, LEGEND MAX, Zombie Aqua, Zombie NIR and Zombie Yellow are the trademarks and property of BioLegend, Inc. ABTS is a registered trademark of Boehringer Mannheim GmbH. BD, BD FACSCalibur, BD FACSCanto, BD FACScan, BD FACSVantage SE, BD FACSAria, BD LSRFortessa, BD CyChrome, BD Influx and BD Horizon are trademarks of Becton, Dickinson and Company. Brilliant Violet, Brilliant Violet 421, BV421, Brilliant Violet 510, BV510, Brilliant Violet 570, BV570, Brilliant Violet 605, BV605, Brilliant Violet 650, BV650, Brilliant Violet 711, BV711, Brilliant Violet 785, BV785 are trademarks of Sirigen Group Ltd. Cascade Blue, Alexa Fluor, Texas Red are registered trademarks and Pacific Blue, Pacific Orange, and Cascade Yellow are trademarks of Molecular Probes, Inc. Coulter and EPICS are registered trademarks and XL, XL-MCL, CyAn, Gallios, and MoFlo are trademarks of Beckman Coulter, Inc. Costar is a registered trademark and Stripwell is a trademark of Corning, Inc. Cy, including Cy3, Cy5, Cy5.5 and Cy7, is a trademark of Amersham Biosciences Ltd. CyAn ADP and MoFlo are trademarks of Dako Denmark A/S. CyTOF is a registered trademark of DVS Sciences. DyLight is a registered trademark of Thermo Fisher Scientific Inc. and its subsidiaries. Trademarks (Continued) FLAG is a registered trademark of Sigma-Aldrich Co. influx is a trademark of Cytopeia, Inc. Synergy and Eclipse are trademarks and Reflection is a registered trademark of icyt Mission Technology, Inc. C6 Flow Cytometer is a registered trademark of Accuri Cytometers, Inc. ImageStream is a registered trademark of Amnis Corporation. MACSQuant is a trademark of Miltenyi Biotec Inc. Patents Alexa Fluor and Pacific Blue : U.S. Patent Nos. 5,696,157; 6,130,101; 5,798,276; 5,830,912; 6,562,632B1; 6,716,979 pending patent applications and corresponding foreign equivalents, owned by Invitrogen Corp. Brilliant Violet : U.S. Patent Nos. 7,914,984; 7,811,755; 7,666,594; 7,629,448; 7,214,489; 7,270,956; 7,144,950; 7,897,684. Licensed from Sirigen, owned by The Regents of the University of California. Allophycocyanin (APC) conjugates: U.S. Patent No. 5,714,386 Cyanine (Cy): U.S. Patent Nos. 4,981,977; 5,268,486 and other patents pending. R-Phycoerythrin (R-PE) and Allophycocyanin (APC): U.S. Patent Nos. 4,542,104 and 5,714,386 PE-Cy7: U.S. Patent No. 4,542,104 APC-Cy7: U.S. Patent No. 5,714,386 Peridinin chlorophyll protein (PerCP): U.S. Patent No. 4,876,190 CD44: U.S. Patent No. 5,108,904 CD62P: U.S. Patent No. 4,783,330 Antibody to Human Fusin (clone 12G5): U.S. Patent No. 5,994,515 12G5: U.S. Patent No. 5,994,515 Antibodies to Human IL-4: U.S. Patent No. 5,041,381 Antibodies to Human and Murine IL-10: U.S. Patent No. 6,239,260 Antibodies to vil-10: U.S. Patent No. 5,756,342 Antibodies to Human and Murine IL-10 Receptor: U.S. Patent No. 5,863,796 Antibodies to Human and Murine IL-17: U.S. Patent No. 6,274,711 Antibodies to Human GM-CSF: U.S. Patent No. 5,070,013 Antibodies to Human and Murine SLAM: U.S. Patent No. 5,576,423 Antibodies to Mouse IL-6 Receptor, Mouse IL-9, Human IL-9: U.S. Patent No. 5,157,112; European Patent No Antibodies to Human IL-9 Receptor: U.S. Patent No. 5,962,269; European Patent No Antibodies to Human and Murine CD317: U.S. Patent No. 7,049,408 Annexin V: U.S. Patent No. 5,834,196 Antibodies to T-bet: U.S. Patent Nos. 7,745,581; 7,393,944; 7,365,169 Antibodies to p27kip1: U.S. Patent Nos. 5,688,665; 6,635,450; 6,242,575; 6,355,774 Antibodies to Human Phospho-ATM (Ser1981): U.S. Patent No. 7,108,992 Antibodies to Gamma-H2A: U.S. Patent Nos. 6,362,317; 6,884,873 Antibodies to Human KIR2DL4: U.S. Patent No. 7,435,801 Antibodies to Human TLR2: U.S. Patent No. 7,071,310B1 and Great Britain Patent No. 9,927,332 Antibodies to Human KIR2DL4: U.S. Patent No. 7,435,801 Antibodies to Mouse TRAIL: Japan Patent No. JP Antibodies to Human and Murine VEGFR-3 (Clone 9D9F9): US Patent Number 6,107,046, EP Patent Number , JP Patent Number , JP Patent Number , CA Patent Number , AU Patent Number Antibodies to ATM Phospho (Ser1981): US Patent Number 7,108,992 Recombinant Human TGF-β1 (carrier-free): U.S. Patent No. 5,104,977, issued April 14, 1992 entitled Purified Transforming Growth Factor Beta, U.S. Patent No. 10/585,499, filed July 6, 2006 entitled Composition and Methods for The High Efficiency Expression of the Transforming Growth Factor-Beta Supergene Family Qdot is a registered trademark of Invitrogen Corp. 9